Biomedical subjects

B Schoner

Publications and source records attributed to B Schoner.

11 recordsLinked to original sources

PubMed1998

Radioimmunoassay of rat leptin: sexual dimorphism reversed from humans.

Adipose tissue secretes leptin, which interacts with receptors in the hypothalamus. In rodent models of obesity, leptin increases metabolism and decreases food intake, which helps to maintain normal body composition. Accurate and precise methods to quantitate circulating leptin concentrations are needed for physiological studies. We developed an RIA to measure leptin in rat plasma, serum, or adipocyte culture fluids. The working range of the assay, defined by the detection limit and the highest calibrator, was 0.5-50 micrograms/L. Recovery of 1.6-11.6 micrograms/L leptin added to serum was 92-103%. The rat leptin RIA correlated well with a previously developed mouse RIA when rat plasma was assayed with both methods (r = 0.94), but the mouse leptin assay underestimated rat leptin in plasma. Within- and between-run CVs were 2.4% to 5.7%. Plasma leptin concentrations correlated directly with percentage of body fat, and correlation improved when the results were separated by gender (r = 0.796, P < 0.001 for males; r = 0.710, P < 0.001 for females). Leptin concentrations were generally higher in male rats than in females; plasma leptin increased 0.60 microgram/L for each percentage of increase in body fat for males but only 0.22 microgram/L for females. We conclude that rat serum/plasma leptin concentrations are accurately and precisely measured with this new RIA.

Adipocytes↗

PubMed1997

Leptin is a four-helix bundle: secondary structure by NMR.

Leptin is a signaling protein that in its mutant forms has been associated with obesity and Type II diabetes. The lack of sequence similarity has precluded analogies based on structural resemblance to known systems. Backbone NMR signals for mouse leptin (13C/15N -labeled) have been assigned and its secondary structure reveals it to be a four-helix bundle cytokine. Helix lengths and disulfide pattern are in agreement with leptin as a member of the short-helix cytokine family. A three-dimensional model was built verifying the mechanical consistency of the identified elements with a short-helix cytokine core.

Amino Acid Sequence↗

PubMed1997

Leptin inhibition of the hypothalamic-pituitary-adrenal axis in response to stress.

Leptin is a newly identified protein hormone that is synthesized and secreted by adipose tissue. Absence of the mature hormone is responsible for the obese phenotype of ob/ob mice. The hypothalamic-pituitary-adrenal axis (HPAA) is activated in ob/ob mice, and chronic administration of leptin to ob/ob mice decreases plasma corticosterone levels, suggesting that the adipose hormone is capable of inhibiting the HPAA. The aim of this study was to determine whether leptin feeds back acutely to inhibit the HPAA of normal mice and rats. Male C57BL mice were injected ip with 100 microl saline and 2 or 4 microg/g BW mouse leptin in saline vehicle, and 4 h later they were subjected to 2 h of restraint stress by taping the hind limbs together or no stress. Hind leg restraint stimulated the HPAA as measured by significant (P < 0.05) elevation of both ACTH and corticosterone. Pretreatment with recombinant mouse leptin blocked the stress-mediated stimulation of both plasma hormones. To determine whether this inhibition was exerted at the hypothalamic level through inhibition of CRH, we studied leptin action on isolated rat hypothalami perifused with Krebs-Ringer buffer containing glucose (5.5 mM). CRH secretion was stimulated by decreasing the glucose concentration of the buffer to 1.1 mM. A surge of CRH was released over a 2-h period (basal integrated release was 14.4 +/- 1.6 pg/2 h, n = 5 and increased to 34.7 +/- 3.1 pg/2 h, n = 14). This response was blocked by mouse leptin in a dose-dependent manner (integrated stimulated CRH secretion was 30.6 +/- 2.5 pg/2 h, n = 5; 20.5 +/- 3.6 pg/2 h, n = 7; 15.3 +/- 4.3 pg/2 h, n = 3 for 1 nM, 3 nM and 30 nM, respectively). Leptin did not alter secretion of ACTH from rat primary cultured pituitary cells. These data demonstrate that leptin can inhibit hypothalamic CRH release, either directly or indirectly through another hypothalamic neuropeptide such as neuropeptide-Y. Dysfunctional leptin, insufficient leptin levels, or leptin resistance should each result in a partial open loop, thereby accounting for elevated glucocorticoid levels that accompany and contribute to many obese phenotypes. Leptin's ability to inhibit CRH release is the likely explanation for its ability to inhibit activation of the HPAA in response to stress.

Adrenocorticotropic Hormone↗

PubMed1992

Sequence similarity between macrolide-resistance determinants and ATP-binding transport proteins.

The three macrolide-resistance-encoding genes, tlrC from Streptomyces fradiae, srmB from Streptomyces ambofaciens, and carA from Streptomyces thermotolerans, encode proteins that possess significant sequence similarity to ATP-dependent transport proteins. The N-terminal and C-terminal halves of these proteins are very similar to each other and contain highly conserved regions that resemble ATP-binding domains typically present within the superfamily of ATP-dependent transport proteins. These observations suggest that the mechanism by which these genes confer resistance to macrolides is due to export of the antibiotics, a process that is driven by energy derived from ATP hydrolysis.

Adenosine Triphosphate↗

PubMed1989

Identification of a gene required for the terminal step in erythromycin A biosynthesis in Saccharopolyspora erythraea (Streptomyces erythreus).

We have identified a transcription unit in the ermE region of the chromosome of the erythromycin (Er)-producing bacterium Saccharopolyspora erythraea (Streptomyces erythreus) that is briefly switched on at about the time that macrolide production commences. Disruption of the transcription unit, herein designated eryG, by insertion of an integrational plasmid vector, caused a block at the terminal step in the biosynthesis of erythromycin, the conversion of erythromycin C to A by O-methylation.

Blotting, Northern↗

PubMed1989

A comparison of two post systems under applied compressive-shear loads.

This study compared a new parallel-tapering, threaded, split-shank post with a well-accepted parallel serrated post under applied compressive-shear loads. The posts were placed in paired, contralateral human teeth to attempt to minimize variation in the tooth model. Both initial and ultimate failure modes were observed, and the clinical significance reported. The Flexi-post system compared favorably with the Para-Post system under the conditions of this research.

Crowns↗

PubMed1987

Vitamin K-dependent carboxylase: possible role of the substrate "propeptide" as an intracellular recognition site.

The liver microsomal vitamin K-dependent carboxylase catalyzes the posttranslational conversion of specific glutamate residues to gamma-carboxyglutamate residues in a limited number of proteins. A number of these proteins have been shown to contain a homologous basic amino acid-rich "propeptide" between the leader sequence and the amino terminus of the mature protein. Plasmids encoding protein C, a vitamin K-dependent protein, containing or lacking a propeptide region were constructed and the protein was expressed in Escherichia coli. The protein products were assayed as substrates in an in vitro vitamin K-dependent carboxylase system. Only proteins containing a propeptide region were substrates for the enzyme. These data support the hypothesis that this sequence of the primary gene product is an important recognition site for this processing enzyme.

Animals↗

PubMed1983

Escherichia coli K-12 F' plasmids carrying insertion sequences IS1 and IS5.

A simple method is described for the detection of the insertion elements IS1 and IS5 in Escherichia coli F' plasmids. Several of these insertion elements are normal constituents of the E. coli chromosome and are located on chromosomal regions carried by the F' plasmids, while several others were probably acquired during the isolation or propagation of the F' plasmids. The F' plasmids carrying copies of IS1 or IS5 have been transferred into Salmonella (a host lacking chromosomal copies of IS1 and IS5) where individual copies can be examined for a variety of properties, including structural similarities and ability to transpose to new sites.

Chromosome Mapping↗

PubMed1983

The nucleotide sequence of the HhaII restriction and modification genes from Haemophilus haemolyticus.

We have determined the nucleotide sequence of a cloned 1710-bp segment of Haemophilus haemolyticus DNA which contains the HhaII restriction (r) and modification (m) genes. The m gene is 513 bp in length and the rgene is 681 bp in length. Both are in the same reading frame, being separated by a 21-bp region. A ribosome-binding site is identified in front of each gene, but no Haemophilus promoter is apparent on the cloned fragment. Transcription originates from a plasmid promotor and proceeds in the direction m to r.

Bacterial Proteins↗

PubMed1981

Distribution of IS5 in bacteria.

Four different strains of Escherichia coli and several other bacteria were examined by Southern analysis for the presence of the insertion element IS5 and IS5-like sequences. Variations in the copy number, degree of homology and restriction pattern of the IS5-like sequence were found among the different organisms. The number and distribution of IS5 sequences do not appear to correlate with the evolutionary relationship of the bacteria in which they occur.

Bacillus subtilis↗

PubMed1981

The nucleotide sequence of IS5 from Escherichia coli.

A 3-kb fragment of Haemophilus haemolyticus DNA which carries the HhaII restriction (r) and modification (m) genes has been cloned into the PstI site of pBR322 (Mann et al., 1978). When propagated in Escherichia coli, it was observed that spontaneous insertions of IS5 inactivated the restriction gene, producing r- mutants at a frequency of 10(-6). Electron microscopy, restriction-site mapping and sequence analysis of two r- plasmids have demonstrated the presence of IS5 at a single target site in both possible orientations. The complete nucleotide sequence of IS5 has been determined. It is 1195 bp long and has inverted terminal repeats of 16 bp. The target site for IS5 in this plasmid is 5'-CTAG. Approx. ten copies of IS5 were found to be present at about the same locations on the E. coli chromosome in various K-12 strains, using Southern hybridization analysis.

Base Sequence↗